In vivo C14-L-alanine metabolism in rat dietary obesity induced by cafeteria diet

The "in vivo" handling of L-alanine in 24 hours starved rats, in which obesity was induced by feeding with cafeteria diet, was compared with that of starved control rats. 14C-alanine was administered in trace amounts in order not to affect the normal handling of this amino acid. The results obtained in blood and liver support a lowered glucose formation from alanine. The specific radioactivities corresponding to lactate, glutamate + glutamine and asparagine as well as total protein and total lipid, were all lowered in the obese group. This strongly suggests that glucose formation from alanine in the liver was impaired. The specific radioactivity of the metabolites studied in the striated muscle are compatible with the above suggestion. It can be concluded that the glucose alanine cycle operation is inhibited in the cafeteria diet starved obese rats.     

Alterations of energy metabolism induced by hexadecane in mice

Two groups of young male OF-1 mice were fed for 60 days with cafeteria or, as controls, with standard pellet diet respectively. At that time, both groups were daily treated with hexadecane (HDK) on the skin. HDK induced a drastic body weight loss much higher in cafeteria than control mice. White adipose tissue were exhausted after 4 days of treatment in controls but not after 10 days in cafeteria ones. HDK resulted in mobilization of liver glycogen in both groups while muscle glycogen decreased slightly in the end. Hexadecane treatment did not result in massively enhanced nitrogen metabolism, as the actual oxidation of amino acids decreased considerably as indicated by the low levels of plasma urea. The results could be explained by powerful and lasting effects of hexadecane on thermogenesis and metabolic reserve balance. The use of this material for pharmacological manipulation of body weight appeared difficult.     

Body weight and tissue composition in rats made obese by a cafeteria diet. Effect of 24 hours starvation

Rat body size and tissue composition changes from pre-weaning to three months age resulted from voluntary hyperphagia triggered by offering a cafeteria diet. The effects of a 24 hour starvation period in both cafeteria and chow fed controls were compared. Obesity develops earlier in females than in males. This difference is related to the growth patterns in both sexes. Obesity occurs at the stages of development when growth rate decreases. Cafeteria fed female rats attained a 32% greater weight than their controls, with lumbar adipose cords that were 4 times heavier and brown interscapular adipose tissue 2 times heavier than controls. The overall cafeteria fed versus chow fed rat differences in the effects of a 24 hour starvation, were minor but less liver glycogen and much more skeletal muscle lipids were mobilized in the cafeteria fed rats than in controls.     

High-energy diets produce different effects on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver in the rat

The influence of feeding rats a high-energy diet for 7 days on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver of the rat was investigated. The incorporation of 3H2O and [U-14C]glucose into fatty acid was measured in vivo. The rats fed the high-energy diets had higher rates of fatty acid synthesis in white adipose tissue than the controls fed on chow, while fatty acid synthesis in brown adipose tissue and liver was either decreased or unchanged relative to that of controls fed on chow. After an oral load of [U-14C]glucose the incorporation of radioactivity into tissue fatty acid was several-fold higher in brown adipose tissue than in white adipose tissue in rats fed on chow. In rats fed the high-energy diets, incorporation of radioactivity into fatty acid in brown adipose tissue was decreased while that into white adipose tissue was either increased (Wistar rats) or unchanged (Lister rats).     

Amino-acid-enzyme activities in brown and white adipose tissues and in the liver of cafeteria rats. Effects of 24 hours starving

Moderate obesity (17% excess body weight) was induced in female rats by offering a "cafeteria" diet during 82 days. The adaptive changes in five amino-acid-metabolism enzymes were determined in liver and white- and brown adipose tissues by comparison with chow fed controls both in the fed and 24-h starved states. Plasma urea levels were lower in the obese and the changes in enzymatic activities pointed to a lower rate of amino-acid metabolism in our dietary obesity group. The levels of activity of amino-acid-metabolism enzymes in brown adipose tissue were higher than in white adipose tissue and in most cases comparable to that of liver. The importance of amino acids as a fuel source in brown adipose tissue thermogenesis cannot be ruled out.     

Thermogenic effects of a "cafeteria" diet on the rat during its reproductive cycle

The heat production and oxygen consumption of intact virgin, pregnant, lactating and postlactating rats has been investigated both in groups fed a "cafeteria" diet as well as in controls. A third group of rats fed the cafeteria diet after parturition has been investigated. Pregnant rats fed a "cafeteria" diet increased their weight faster than controls. During lactation no increases in weight were observed, and in postlactation both "cafeteria" groups attained the same values higher than controls. The ingestion of a "cafeteria" diet resulted in higher heat production in all groups except lactating rats, which--in addition--showed higher heat outputs than all the other groups when the actual data were corrected by metabolic weight according to the surface law. The high lactation heat production (and oxygen consumption) can be a consequence of increased metabolic activity in the rat organism, devoted to milk production. It can be concluded that during lactation the dam energy output through the milk must absorb any increases in energy input due to the more densely-packed energy in the "cafeteria" diet, and this did not result in increased heat production.     

Triglyceride Uptake in Muscles in Rats

Exogenous lipid is assimilated with different priorities in adipose tissue regions and varies in the fasting and fed conditions. The quantitative role of uptake of lipid in muscle has not been evaluated. In order to examine the uptake in other than adipose tissues, U¹⁴C-oleic acid in sesame oil was administered orally to conscious rats, and lipid label measured after different times in serum, heart, liver, mesenteric, retroperitoneal, inguinal and epididymal fat pads, as well as in red and white parts of gastrocnemius, extensor digitorum longus and soleus muscles. Lipid uptake in total adipose tissue was calculated from dissected adipose tissues plus lipids extracted from the eviscerated, skinned carcass. Lipid uptake in total muscle tissue was estimated from label in dissected muscles plus that in the carcass, assuming similar intracellular lipid contents and radioactivity as that averaged from dissected muscles. Lipid uptake in the liver was calculated from directly extracted lipid. Four hours after lipid administration to fed rats lipid radioactivity in heart and serum was minimal and had essentially disappeared at 8 hours. Liver label declined rapidly from peak values at or before 4 hours. Adipose tissue radioactivity increased gradually up to 16 hours and then decreased. Label in muscles was highest at 4 hours in the red gastrocnemius, and then decreased, while the other muscles showed a constant radioactivity over the observation period (24 hours). Radioactivity expressed per unit muscle mass seemed to be proportional to the oxidative capacity of muscles. In comparisons between fed and fasted rats at 16 hours, when adipose tissue label peaked, liver, individual muscles and carcass did not show any significant differences while adipose tissue label was fivefold higher in fed than fasted rats. The distribution of total measured lipid radioactivity between total adipose tissue, total muscle tissue and liver in fed rats at this time-point was 76. 8, 14. 4 and 8. 8% respectively, and in the fasted state 26. 4, 51. 6 and 22. 0%. These estimations suggest that lipid uptake in the fed state is dominated by adipose tissue, while in the fasted state the lipid uptake is higher in muscles than adipose tissues. It was concluded that uptake of absorbed, exogenous triglyceride in muscle is of significance, particularly in the fasted state. This lipid has a half life of several days. It is suggested that this lipid is oxidized in situ, contributing with a hidden fraction to lipid energy needs, or partially transferred to adipose tissue. Lipid uptake in muscle probably constitutes a significant fraction of assimilated exogenous lipid, particularly in the fasting state.     

Adaptation of adenosylmethionine metabolism and methionine recycling to variations in dietary methionine in the rat

Weanling rats were fed a casein-based diet supplemented to give dietary methionine (Met) concentrations of 0.41, 0.61, and 1.50%. After 2 weeks of feeding, the rats received intraperitoneally 800 nCi of 2-14C-labeled and/or methyl-3H-labeled L-Met. The animals were killed 20 min, 1 hr, or 2 hr after the isotope injection and the specific radioactivity of adenosylmethionine (AdoMet) as well as the total acid-soluble radioactivity was analyzed in the liver and skeletal muscle. Met concentrations of the liver and skeletal muscle were increased 20-fold by the diet containing 1.50% of Met. In the liver, but not in skeletal muscle, accumulation of AdoMet closely followed changes in Met concentration. Within 2 hr after intraperitoneal injection, the rate of disappearance of 3H label from the acid-soluble fraction was slow in both tissues; increasing in the liver and decreasing in skeletal muscle with increasing dietary Met concentration. At the same time, disappearance of 14C label was slow in both tissues in the rats fed the toxic Met diet, and also in the liver of the rats fed the Met-deficient diet. Decline of the specific radioactivity of the AdoMet pool with respect to 3H label was similar to that of 14C label in the skeletal muscle at all dietary Met concentrations. In the liver, the rate of disappearance of 14C label from the AdoMet pool was markedly increased and that of the 3H label slightly decreased with increasing dietary Met supply. Met deprivation resulted in rapid disappearance of 3H label from the hepatic AdoMet pool, whereas the disappearance of the 14C label was very slow. The results indicate that hepatic Met recycling is very effective with deficient or adequate dietary Met concentrations. In skeletal muscle, the capacity to catabolize extra Met is very limited and continuous flow of Met to liver takes place. Unlike in the liver, in skeletal muscle the transsulfuration route is not adaptable to changes in Met supply and plays a minor role in Met catabolism. The approach used to determine the efficacy and adaptation of methionine salvage pathways by following simultaneously the decline of the specific radioactivities of the methyl group and the methionyl carbon chain of AdoMet following intraperitoneal injection of double-labeled Met has several advantages over that used in literature reports. It offers a reliable means of observing these metabolic pathways in whole animals without disruption of metabolite fluxes.     

Effects Of A Post-Weaning Cafeteria Diet In Young Rats: Metabolic Syndrome,Reduced Activity And Low Anxiety-Like Behaviour

Among adolescents, overweight, obesity and metabolic syndrome are rapidly increasing in recent years as a consequence of unhealthy palatable diets. Animal models of diet-induced obesity have been developed, but little is known about the behavioural patterns produced by the consumption of such diets. The aim of the present study was to determine the behavioural and biochemical effects of a cafeteria diet fed to juvenile male and female rats, as well as to evaluate the possible recovery from these effects by administering standard feeding during the last week of the study. Two groups of male and female rats were fed with either a standard chow diet (ST) or a cafeteria (CAF) diet from weaning and for 8 weeks. A third group of males (CAF withdrawal) was fed with the CAF diet for 7 weeks and the ST in the 8th week. Both males and females developed metabolic syndrome as a consequence of the CAF feeding, showing overweight, higher adiposity and liver weight, increased plasma levels of glucose, insulin and triglycerides, as well as insulin resistance, in comparison with their respective controls. The CAF diet reduced motor activity in all behavioural tests, enhanced exploration, reduced anxiety-like behaviour and increased social interaction; this last effect was more pronounced in females than in males. When compared to animals only fed with a CAF diet, CAF withdrawal increased anxiety in the open field, slightly decreased body weight, and completely recovered the liver weight, insulin sensitivity and the standard levels of glucose, insulin and triglycerides in plasma. In conclusion, a CAF diet fed to young animals for 8 weeks induced obesity and metabolic syndrome, and produced robust behavioural changes in young adult rats, whereas CAF withdrawal in the last week modestly increased anxiety, reversed the metabolic alterations and partially reduced overweight.     

Effects of starvation and a high-energy diet on rat blood compartmentation of injected radioactive alanine and glucose

The effects of starvation upon blood compartmentation and differential handling of glucose and alanine have been studied in control and cafeteria-fed rats. The injection of radioactive glucose resulted in higher specific radioactivities in intracellular glucose but lower intracellular amino acid specific radioactivities when compared with the plasma values. The cells/plasma specific radioactivity ratios increased dramatically in cafeteria rats with starvation. The injection of radioactive alanine resulted in higher cell than plasma glucose specific activities, and lower cell amino acid specific activities. All these parameters increased after a 24-hours starvation period. It is concluded that glucose synthetised by the liver is released mainly into the blood intracellular pool, being later liberated into the plasma or directly into the tissues.     

Plasma amino acids of lean and obese Zucker rats subjected to a cafeteria diet after weaning.

Plasma amino acids of Zucker obese (fa/fa) and lean (Fa/?) rats fed either a reference nonpurified pellet or a cafeteria diet have been studied from 30 to 60 days after birth. Obese rats showed higher plasma branched chain amino acid levels but similar total amino acids, urea and glucose concentrations. The ingestion of a cafeteria diet induced higher levels in many amino acids, as well as in the composite figure in lean rats, but failed to alter total 2-amino nitrogen concentrations in obese rats, despite high levels in several non-essential amino acids and lower values in essential amino acids; urea levels were much lower in rats fed the cafeteria diet. The results are consistent with an impairment of amino acid nitrogen elimination via urea cycle in cafeteria diet-fed rats. This is independent of the hyperinsulinemia-driven plasma accumulation of several essential amino acids induced by genetic obesity. The effects were, then additive.     

Morphofunctional changes in gastrointestinal tract of rats due to cafeteria diet.

Female rats fed a cafeteria diet from birth developed obesity at 60 days of age and their stomach, small intestine and caecum were enlarged when compared with controls, i.e. these regions had greater food storage capacity. In spite of the enlargement, these regions had similar or reduced weight and linear density, which is seen as proof of reduced mechanical performances. Cafeteria diet produced increased glucose duodenal absorption in older animals unlike the typical reduction known in controls. Tryptophan absorption was maintained high in adulthood, compensating for the low structural nutritive properties of the cafeteria diet. The results are interpreted as an adaptation to the cafeteria diet effects and properties: the characteristic overeating of foodstuffs with greater energy density, lower mechanical requirements and lower structural nutritive value than pelleted chow.     

Uptake of radiolabeled copper from portal blood containing fructose or glucose

The present investigation was designed to study the uptake of⁶⁷Cu when administered directly, into the portal vein, along with either functose or glucose, by the liver and extrahepatic tissues. Following weaning, male Sprague-Dawley rats were fed for 3 wk either commercial laboratory ration (chow) or semipurified diets deficient in Cu (0.6 ppm) or supplemented with Cu (6.0 ppm) and containing 62% carbohydrate as either fructuse or cornstarch. After an overnight fast, a single dose of rat plasma (0.1 mL) containing fructose or glucose extrinsically labeled with⁶⁷Cu was injected directly into their portal vein. Although not always statistically significant, rats fed chow retained more radioactivity in the liver and several extrahepatic tissues when⁶⁷Cu was administered with fructose than with glucose. Regardless of Cu status, rats fed diets containing fructose retained more radioactivity in extrahepatic tissues than rats fed starch. There was an increased uptake of⁶⁷Cu by the liver, blood, muscle, and fat pad when fructose as compared to glucose was injected in combination with the isotope. These data strongly suggest that Cu requirements or utilization are greater when fructose is the main dietary carbohydrate. The results may also in part explain the reason for the increased severity of Cu deficiency in rats fed fructose.     

Sucrose feeding effects inhibition of gamma-glutamyltranspeptidase in the liver of the rat: possible mediation by thyroid hormone

The effect of sucrose on Fischer 344 rat liver gamma-glutamyltranspeptidase (gammaGT) was studied: in adults fed sucrose for 3 weeks; and rats exposed to sucrose from the 18th day of gestation to the 40th day after birth. Rats fed regular rodent chow served as controls. Sucrose caused mild lipemia; and in the liver an increase in size and fat build-up without damage. In adult sucrose-fed rats, compared to controls, plasma glucose levels were increased: 1.12-, 1.40- and 1.13-fold after 24, 48h and 3 week consumption of sucrose, respectively. Insulin levels were unaltered for the first week of sucrose consumption but increased from control levels: 16% at 1 week, and 2.0-fold at 3 weeks. The T3 levels were comparable to control levels 24h after the sucrose was started and were increased: 1.22-, 1.13- and 1.12-fold at 48h, 1 and 3 weeks, respectively. The T4 levels were comparable at all time points between sucrose-fed and control rats. Liver gammaGT activity exhibited a steady decrease from control levels: after 24, 48h, 1 and 3 weeks of sucrose feeding the decrease was 5, 8, 21 and 37%, respectively in homogenates; and 10, 17, 24 and 41%, respectively in plasma membranes. Perinatal sucrose exposure effected in 40-day-old rats, compared controls: a 1.09-fold increase in plasma glucose; no change in plasma insulin; an increase of 1.15- and 1.39-fold in plasma levels of total and free T3, respectively; a decrease of 20 and 14% in plasma levels of total and free T4, respectively. gammaGT activity was decreased in liver plasma membranes isolated from sucrose-exposed rats relative to those of control: 80% in the male; 82% in the female. Relative specific activities of gammaGT were the same in both males: 15.4 and 16.1 in control and sucrose-exposed male rats, respectively; and females: 14.1 and 15.4 in control and sucrose-exposed female rats, respectively. gammaGT was 2-fold higher in the livers of female relative to male rats in sucrose-exposed and control groups. Kidney gammaGT activities were the same in control and sucrose-exposed rats. The involvement of T3 in the sucrose-induced decrease in liver gammaGT is discussed.     

Nitrogen balance discrepancy in Wistar rats fed a cafeteria diet.

The nitrogen balance of Wistar rats aged 30-45 and 45-60 days fed either control or cafeteria diet has been determined by measuring the intake fecal and urinary excretion and nitrogen deposition in the body. The efficiency of extraction of dietary nitrogen was higher for cafeteria diet-fed rats, which showed a lower nitrogen excretion and higher body nitrogen accretion than controls. The accurate measurement of nitrogen intake, excretion and deposition showed a consistent proportion of nitrogen unaccounted for (10-26% of net intake) in the studied fractions, which proportion was higher in the youngest cafeteria diet-fed rats.     

Glutathione depletion by hyperphagia-induced obesity

Reduced glutathione (GSH) levels in freeze-clamped livers of rats and mice in which hyperphagia is induced by cafeteria diet are 45% lower than in controls. Freshly isolated hepatocytes from mice fed cafeteria diet show a 45% decrease in GSH concentration and a 54% decrease in oxidized glutathione (GSSG) concentration when compared with controls. The rate of GSH synthesis in isolated hepatocytes from control mice is significantly higher than in those from mice fed cafeteria diet. Oral GSH is effective to prevent the decrease in hepatic GSH levels found in cafeteria fed mice.     

Metabolic utilization of muscular L-proline in 24-hr starved rats.

1. The aim of this paper was to study the in vivo skeletal muscle L-proline related to its destination to other key tissues such as liver and intestine as well as to give some insight into the role of blood cells in proline handling. 2. L-U-[14C]Proline was injected intramuscularly and following by sampling of blood, liver, intestine and contralateral muscle at 20 and 30 min after injection. 3. The distribution of radioactivity between blood cells and plasma and in total and individual amino acids, protein and glycogen fractions was determined in the above tissues. 4. The pattern of well fed rats was compared with those submitted to 24-hr complete starvation. 5. During starvation a minor degree of proline oxidation occurs. 6. The main destruction of proline in the liver seem to be the synthesis of proteins. 7. The radioactivity recovered in the blood proline fraction of starved rats is twice that of the fed rats and that it could be attributed mainly to plasma protein. 8. We have obtained in vivo evidence for the role of erythrocyte in the interorgan proline transport.     

Studies on lipogenesis in vivo. Effect of dietary fat or starvation on conversion of [14C]glucose into fat and turnover of newly synthesized fat

1. Lipogenesis was studied in vivo by giving mice 250mg. meals of [U-(14)C]glucose and measuring the disposition and incorporation of label. About 48% of the (14)C dose was eliminated as (14)CO(2) in the first 2hr. At 60min. after administration, 1.0, 1.9 and 11.9% of the dose was recovered as liver glycogen, liver fatty acid and carcass fatty acid respectively. Of the [(14)C]glucose converted into fat in the epididymal pads about 90% was present as glyceride fatty acid and 10% as glyceride glycerol. 2. Hepatic synthesis of fatty acid was depressed by dietary fat to a much greater extent than was synthesis outside the liver. Both feeding with fat and starvation decreased the proportion of the label taken up by adipose tissue present as fat (triglyceride) and increased the proportion of triglyceride label present as glyceride glycerol. These results are consistent with the hypothesis that the primary action of both these conditions in decreasing fat synthesis is to inhibit synthesis of fatty acids. 3. Turnover of body fat labelled in vivo from [U-(14)C]glucose was estimated from the decline in radioactivity measured over the first 24hr. of the experiment. The half-life of liver and extrahepatic fatty acids (excluding epididymal fat) was 16hr. and 3 days respectively. In contrast, no measurable decrease in radioactivity of the fatty acids of epididymal fat was observed for 7 days after administration of the [U-(14)C]glucose.     

Positive correlation of skeletal muscle UCP3 mRNA levels with overweight in male,but not in female,rats

The objective of this study was to investigate the sex-dependent regulation of skeletal muscle uncoupling protein (UCP)3 mRNA expression in response to overweight and its relationship with serum levels of free fatty acids, leptin, and insulin. Two obesity models were used: rats made obese by feeding them with a cafeteria diet for 14 wk, and postcafeteria overweight rats fed a chow diet for 10 wk after consuming the cafeteria diet for 14 wk. The effects of 24-h fasting were studied in postcafeteria rats and their age-matched controls. The cafeteria rats ate a high-fat diet and attained an excess body weight that was higher in females (+59%) than in males (+39%). A trend to higher induction of abdominal muscle UCP3 mRNA in male rats than in females after cafeteria diet was apparent (+116% increase vs. +26% increase). Postcafeteria male but not female rats still showed the tendency to have increased UCP3 mRNA levels relative to their age-matched controls. A linear regression analysis showed a significant positive correlation of the UCP3 mRNA levels with overweight and with serum levels of leptin and insulin in males, but not in females, and no correlation with serum free fatty acid levels. A subsequent correlation analysis and a multiple linear regression analysis showed that overweight was the only parameter actually related to UCP3 mRNA levels in males. Fasting-induced upregulation of muscle UCP3 mRNA levels was higher in males (5- to 7-fold) than in females (3- to 4-fold). Our results point to the existence of sex-associated differences in the control of muscle UCP3 expression in response to overweight and fasting, with an impaired induction in female rats under both conditions. The correlation of abdominal muscle UCP3 mRNA expression with overweight in males could be related to their relative resistance to gain weight after chronic overeating of a cafeteria diet, by the purported role of UCP3 in the regulation of lipid utilization.     

The effects of elk velvet antler consumption on the rat: development, behavior, toxicity and the activity of liver gamma-glutamyltranspeptidase

The effect of exposure to, followed by consumption of, a diet containing 10% powdered elk velvet antler (EVA) from the 18th day of gestation to the 88th day after birth was examined in male and female Fischer 344 rats. There were no teratogenic effects of EVA exposure in utero or differences in birth outcomes between pups born to regular chow fed and EVA chow fed dams. Growth curves of the EVA fed rats were identical to those of regular chow fed rats, as were developmental milestones of pinna development and eye-opening. Acoustical startle and righting reflexes, developmental and behavioral indices, were identical. Blood glucose levels were comparable in EVA chow fed and regular chow fed rats, indicating that EVA is without effect on glucose balance. There were no signs of toxicity in the EVA chow fed compared to regular chow fed rats as judged from plasma enzyme markers of liver damage: plasma levels of alanine aminotransferase were 50% lower in EVA chow fed rats compared to regular chow fed rats; and plasma levels of gamma-glutamyltranspeptidase (gammaGT) were the same. The activity of gammaGT displayed a decrease in the livers of EVA chow fed rats, more so in the male (22%) than in the female (14%), suggestive of an androgenic effect. A possible hepatobeneficial effect of the EVA induced decrease in liver gammaGT is discussed. In summary, dietary10% EVA chow is without long term effect on growth, development and behavior is non-toxic and may be hepatobeneficial.